HA modified delivery systems will bind to any cell that possesses CD44, as recently shown for macrophages . Finally, CS modification of CD44 (which occurs in melanoma) negatively regulates HA binding [31, 32]. Figure 1 Schematic structure of CD44. The hyaluronate/hyaluronic acid (HA) binding site is in the N-terminal portion (Link module) of CD44 (residues Arg41-Tyr105) [33–35], while the CS modification primarily occurs at Ser180 Inhibitors,research,lifescience,medical . The alternatively spliced … In addition to binding to HA, CS modified CD44 binds collagen [42–44]. The sequence to which CD44 binds within the type IV collagen
triple helix has been identified as α1(IV)1263–1277 (gene-derived sequence Gly-Val-Lys-Gly-Asp-Lys-Gly-Asn-Pro-Gly-Trp-Pro-Gly-Ala-Pro) Inhibitors,research,lifescience,medical [41, 45]. Efficient binding is dependent upon CS modification of CD44 . This sequence is not
bound by collagen-binding integrins [41, 46]. We have previously constructed α1(IV)1263–1277 based triple-helical “peptide-amphiphiles” (PAs) [general structure Cn-(Selleckchem CHIR 99021 Gly-Pro-Hyp)4-Gly-Val-Lys-Gly-Asp-Lys-Gly-Asn-Pro-Gly-Trp-Pro-Gly-Ala-Pro-(Gly-Pro-Hyp)4-NH2] specific for CD44/CSPG [41, 47–49]. M14#5 human melanoma cells Inhibitors,research,lifescience,medical bound to C14, C16, or C18α1(IV)1263–1277PA with EC50 approximately 0.08–0.5μM [41, 46, 50]. The amphiphilic design of the PA construct facilitates the anchoring of the functional “head group” of the construct to the liposome surface by the insertion of the hydrophobic acyl “tail” into the lipid bilayer. This in turn allows the hydrophilic head group or targeting the portion of the PA to protrude outward from the liposomal Inhibitors,research,lifescience,medical surface making it available to interact with the CD44/CSPG receptor. The incorporation of the α1(IV)1263–1277PAs into rhodamine-loaded liposomes did not destabilize these systems and conferred Inhibitors,research,lifescience,medical targeting selectivity to liposomes against
cell lines varying in the CD44 expression based on the receptor/PA ligand recognition . In the current study we evaluated the stability of distearoyl phosphatidylglycerol-(DSPG-)distearoyl phosphatidylcholine (DSPC) DOX-loaded liposomes both with and without the α1(IV)1263–1277PA. We incorporated PEG-2000 into the liposomal systems to allow for increased circulation Adenylyl cyclase times in vivo [51–54]. The efficacies of the various liposomal nanoDDSs were evaluated by quantifying their cytotoxic effects against cell lines with varying levels of CD44/CSPG expression (Scheme 1) and in a B16F10 mouse melanoma model system. Scheme 1 Schematic depiction of targeted liposomal delivery to CD44/CSPG metastatic melanoma cells. The α1(IV)1263–1277PA (red alkyl tail and green peptide head group) is incorporated into liposomes along with DOX (blue circles). The liposome … 2. Materials and Methods 2.1. Chemicals All phospholipids (Cat# 850365, 840465, and 880120) and cholesterol (Cat# 700000) were purchased from Avanti Polar Lipids.